Oocyte-specific gene knockdown by intronic artificial microRNAs driven by Zp3 transcription in mice

• Published in: Journal of Reproduction and Development Vol. 67; no. 3; pp. 229 - 234
• Main Authors: SASAKI, Keisuke, TAKAOKA, Saaya, OBATA, Yayoi
• Format: Journal Article
• Language: English
• Published: Japan The Society for Reproduction and Development 01.01.2021; Japan Science and Technology Agency
• Subjects: Artificial microRNA; Genes; Genetic screening; Genome editing; Green fluorescent protein; MicroRNAs; miRNA; Oocyte; Oocytes; Rodents; Somatic cells; Technology Report; Transcription; Transgenic; Transgenic mice; Zona pellucida glycoprotein 3; Transgenic; Genome editing; Artificial microRNA; Zona pellucida glycoprotein; Oocyte
• ISSN: 0916-8818; 1348-4400
• DOI: 10.1262/jrd.2020-146

Conditional knockout technology is a powerful tool for investigating the spatiotemporal functions of target genes. However, generation of conditional knockout mice involves complicated breeding programs and considerable time. A recent study has shown that artificially designed microRNAs (amiRNAs), inserted into an intron of the constitutively expressed gene, induce knockdown of the targeted gene in mice, thus creating a simpler method to analyze the functions of target genes in oocytes. Here, to establish an oocyte-specific knockdown system, amiRNA sequences against enhanced green fluorescent protein (EGFP) were knocked into the intronic sites of the Zp3 gene. Knock-in mice were then bred with EGFP transgenic mice. Our results showed that Zp3-derived amiRNA successfully reduced EGFP fluorescence in the oocytes in a size-dependent manner. Importantly, knockdown of EGFP did not occur in somatic cells. Thus, we present our knockdown system as a tool for screening gene functions in mouse oocytes.